Molecular Cloning: A Laboratory Manual, Book 3 |
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Page 17-22
A Laboratory Manual Joseph Sambrook, Tom Maniatis. ENGINEERING A RESTRICTION SITE Place a restriction site ... enzyme leaves a 5 ' overhang upon digestion . Site 2 must lie between site 1 and the start of the gene , and it must be closer to ...
A Laboratory Manual Joseph Sambrook, Tom Maniatis. ENGINEERING A RESTRICTION SITE Place a restriction site ... enzyme leaves a 5 ' overhang upon digestion . Site 2 must lie between site 1 and the start of the gene , and it must be closer to ...
Page F-6
... restriction enzyme ( s ) . The amount of foreign DNA digested should be sufficient to yield approxi- mately 0.2 μg of the target fragment . The digestion should be carried out in a volume of 20 μl or less . In a separate tube , digest ...
... restriction enzyme ( s ) . The amount of foreign DNA digested should be sufficient to yield approxi- mately 0.2 μg of the target fragment . The digestion should be carried out in a volume of 20 μl or less . In a separate tube , digest ...
Page 1-38
... Restriction and modification systems dam methylase , 2.57 , 5.15 dcm methylase , 5.15 deficient strains for growing ... enzyme ( s ) , 5.3 , 5.32 . See also Restriction and modification systems ; Restriction mapping of foreign DNA ...
... Restriction and modification systems dam methylase , 2.57 , 5.15 dcm methylase , 5.15 deficient strains for growing ... enzyme ( s ) , 5.3 , 5.32 . See also Restriction and modification systems ; Restriction mapping of foreign DNA ...
Contents
Selectable Markers 1 | 16-7 |
VECTOR SYSTEMS 16 | 16-17 |
DEVELOPMENT OF PLASMID CLONING VECTORS | 16-24 |
Copyright | |
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Acad acetate acrylamide activity Adjust the volume agarose aliquots amount antibody antigen antisera aqueous assay autoclaving bacterial bacteriophage T4 BamHI Biol calcium cDNA cell lines centrifugation chloramphenicol cloned gene column containing culture deionized H₂O dhfr dilution Dissolve dithiothreitol DNA polymerase EDTA EDTA pH 8.0 efficiency elution encoding Escherichia coli ethanol ethidium bromide eukaryotic extract fragment glycerol hybridization immunoglobulin immunoprecipitation Incubate ligation lysate mammalian cells medium method mg/ml microfuge tube minutes at 4°C minutes at room mixture mRNAs NaCl Natl nitrocellulose nitrocellulose filter nucleic acid pellet phosphate plasmid plate PMSF polyacrylamide polyclonal precipitate prepared promoter protoplasts radioactive radiolabeled reaction reagent recombinant remove restriction enzyme Resuspend room temperature sample SDS gel-loading buffer SDS-polyacrylamide gel sequences serum sodium sterile stock solution stored strain supernatant target protein Telephone termini transcription transfection transfer transient expression Tris Cl pH wash western blotting µg/ml